1. Field of the Invention
The present invention relates to methods and compositions for treating diseases in a subject associated with one or more self-protein(s), -polypeptide(s) or -peptide(s) that are present in the subject and involved in a non-physiological state. The present invention also relates to methods and compositions for preventing diseases in a subject associated with one or more self-protein(s), -polypeptide(s) or -peptide(s) that are present in the subject and involved in a non-physiological state. The invention further relates to the identification of a self-protein(s), -polypeptide(s) or -peptide(s) present in a non-physiological state and associated with a disease. The invention also relates to the administration of a polynucleotide encoding a self-protein(s), -polypeptide(s) or -peptide(s) present in a non-physiological state and associated with a disease. The invention also relates to modulating an immune response to a self-protein(s), -polypeptide(s) or -peptide(s) present in an animal and involved in a non-physiological state and associated with a disease. The invention is more particularly related to the methods and compositions for treating or preventing autoimmune diseases associated with one or more self-protein(s), -polypeptide(s) or -peptide(s) present in the animal in a non-physiological state such as in multiple sclerosis, rheumatoid arthritis, insulin dependent diabetes mellitus, autoimmune uveitis, primary biliary cirrhosis, myasthenia gravis, Sjogren's syndrome, pemphigus vulgaris, scleroderma, pernicious anemia, systemic lupus erythematosus (SLE) and Grave's disease. The invention is also particularly related to the methods and compositions for treating or preventing neurodegenerative diseases associated with one or more self-protein(s), -polypeptide(s) or -peptide(s) present in the animal in a non-physiological state such as Alzheimer's disease, Parkinson's disease, Huntington's disease, and transmissable spongiform encephalopathy (prion disease with the most common form referred to as Creutzfeldt-Jakob disease). The invention is further particularly related to other diseases associated with one or more self-protein(s), -polypeptide(s) or -peptide(s) present in the animal in a non-physiological state such as osteoarthritis, spinal cord injury, obesity, hypertension, peptic ulcer disease, depression, gout, migraine headaches, hyperlipidemia and coronary artery disease. The invention is further particularly related to disease(s) such as disseminated encephalomyelitis associated with one or more self-proteins(s), -polypeptides arising out of the administration of, for example, smallpox vaccine. The invention is also related to the means and methods for treating or preventing disease associated with self-protein(s), -polypeptide(s), or -peptide(s) that are present in an animal that is in a non-physiological state. The invention is further related to the treatment of animals comprising the administration of a polynucleotide encoding self-protein(s), -polypeptide(s), or -peptide(s) that are present non-physiologically or involved in a non-physiologic process in the animal.
2. Autoimmune Disease and Modulation of the Immune Response
Autoimmune disease is any disease caused by adaptive immunity that becomes misdirected at healthy cells and/or tissues of the body. Autoimmune disease affects 3% of the U.S. population and likely a similar percentage of the industrialized world population. (Jacobson et al., Clin Immunol Immunopathol 84, 223-43, 1997). Autoimmune diseases are characterized by T and B lymphocytes that aberrantly target self-proteins, -polypeptides, -peptides, and/or other self-molecules causing injury and or malfunction of an organ, tissue, or cell-type within the body (for example, pancreas, brain, thyroid or gastrointestinal tract) to cause the clinical manifestations of the disease (Marrack et al., Nat Med 7, 899-905, 2001). Autoimmune diseases include diseases that affect specific tissues as well as diseases that can affect multiple tissues. This may, in part, for some diseases depend on whether the autoimmune responses are directed to an antigen confined to a particular tissue or to an antigen that is widely distributed in the body. The characteristic feature of tissue-specific autoimmunity is the selective targeting of a single tissue or individual cell type. Nevertheless, certain autoimmune diseases that target ubiquitous self-proteins can also affect specific tissues. For example, in polymyositis the autoimmune response targets the ubiquitous protein histidyl-tRNA synthetase, yet the clinical manifestations primarily involved are autoimmune destruction of muscle.
The immune system employs a highly complex mechanism designed to generate responses to protect mammals against a variety of foreign pathogens while at the same time preventing responses against self-antigens. In addition to deciding whether to respond (antigen specificity), the immune system must also choose appropriate effector functions to deal with each pathogen (effector specificity). A cell critical in mediating and regulating these effector functions is the CD4+ T cell. Furthermore, it is the elaboration of specific cytokines from CD4+ T cells that appears to be the major mechanism by which T cells mediate their functions. Thus, characterizing the types of cytokines made by CD4+ T cells as well as how their secretion is controlled is extremely important in understanding how the immune response is regulated.
The characterization of cytokine production from long-term mouse CD4+ T cell clones was first published more than 10 years ago (Mosmann et al., J. Immunol. 136:2348-2357, 1986). In these studies, it was shown that CD4+ T cells produced two distinct patterns of cytokine production, which were designated T helper 1 (Th1) and T helper 2 (Th2). Th1 cells were found to predominantly produce interleukin-2 (IL-2), interferon-γ (IFN-γ) and lymphotoxin (LT), while Th2 clones predominantly produced IL-4, IL-5, IL-6, and IL-13 (Cherwinski et al., J. Exp. Med. 169:1229-1244, 1987). Somewhat later, additional cytokines, IL-9 and IL-10, were isolated from Th2 clones (Van Snick et al:, J. Exp: Med. 169:363-368, 1989) (Fiorentino et al., J. Exp. Med. 170:2081-2095, 1989). Finally, additional cytokines, such as IL-3, granulocyte macrophage colony-stimulating factor (GM-CSF), and tumor necrosis factor-α (TNF-α) were found to be secreted by both Th1 and Th2 cells.
Autoimmune disease encompasses a wide spectrum of diseases that can affect many different organs and tissues within the body as outlined in the table above. (See e.g. Paul, W. E. (1999) Fundamental Immunology, Fourth Edition, Lippincott-Raven, New York.)
TABLE IPrimary Organ(s) TargetedDiseasethyroidHashimoto's DiseasethyroidPrimary myxodaemathyroidThyrotoxicosisstomachPernicious anemiastomachAtrophic gastritisadrenal glandsAddison's diseasepancreatic isletsInsulin dependent diabetes mellituskidneysGoodpasture's syndromeneuromuscular junctionMyasthenia gravisleydig cellsMale infertilityskinPemphigus vulgarisskinPemphioideyesSympathetic ophthalmiaeyesPhacogenic uveitisbrainMultiple sclerosisred blood cellsHemolytic anemiaplateletsIdiopathic thrombocytopenic purpurawhite blood cellsIdiopathic leucopeniabiliary treePrimary biliary cirrhosisbowelUlcerative colitisarteriesAtherosclerosissalivary and lacrimal glandsSjogren's syndromesynovial jointsRheumatoid arthritismusclePolymyositismuscle and skinDermatomyositisskinSclerodermaskin, joints, muscle, blood cellsMixed connective tissue diseaseclotting factorsAnti-phospholipid diseaseskinDiscoid lupus erythematosusskin, joints, kidneys, brain, bloodSystemic lupus erythematosus (SLE)cells
Current therapies for human autoimmune disease, include glucocorticoids, cytotoxic agents, and recently developed biological therapeutics. In general, the management of human systemic autoimmune disease is empirical and unsatisfactory. For the most part, broadly immunosuppressive drugs, such as corticosteroids, are used in a wide variety of severe autoimmune and inflammatory disorders. In addition to corticosteroids, other immunosuppressive agents are used in management of the systemic autoimmune diseases. Cyclophosphamide is an alkylating agent that causes profound depletion of both T- and B-lymphocytes and impairment of cell-mediated immunity. Cyclosporine, tacrolimus, and mycophenolate mofetil are natural products with specific properties of T-lymphocyte suppression, and they have been used to treat SLE, RA and, to a limited extent, in vasculitis and myositis. These drugs are associated with significant renal toxicity. Methotrexate is also used as a “second line” agent in RA, with the goal of reducing disease progression. It is also used in polymyositis and other connective-tissue diseases. Other approaches that have been tried include monoclonal antibodies intended to block the action of cytokines or to deplete lymphocytes. (Fox, D. A. Am. J. Med; 99:82-88 1995). Treatments for multiple sclerosis (MS) include interferon β and copolymer 1, which reduce relapse rate by 20-30% and only have a modest impact on disease progression. MS is also treated with immunosuppressive agents including methylprednisolone, other steroids, methotrexate, cladribine and cyclophosphamide. These immunosuppressive agents have minimal efficacy in treating MS. Current therapy for rheumatoid arthritis (RA) utilizes agents that non-specifically suppress or modulate immune function such as methotrexate, sulfasalazine, hydroxychloroquine, leuflonamide, prednisone, as well as the recently developed TNFα antagonists etanercept and infliximab (Moreland et al., J Rheumatol 28, 1431-52., 2001). Etanercept and infliximab globally block TNFα, making patients more susceptible to death from sepsis, aggravation of chronic mycobacterial infections, and development of demyelinating events.
In the case of organ-specific autoimmunity, a number of different therapeutic approaches have been tried. Soluble protein antigens have been administered systemically to inhibit the subsequent immune response to that antigen. Such therapies include delivery of myelin basic protein, its dominant peptide, or a mixture of myelin proteins to animals with experimental autoimmune encephalomyelitis and humans with multiple sclerosis (Brocke et al., Nature 379, 343-6, 1996; Critchfield et al., Science 263, 1139-43., 1994; Weiner et al., Annu Rev Immunol 12, 809-37, 1994), administration of type II collagen or a mixture of collagen proteins to animals with collagen-induced arthritis and humans with rheumatoid arthritis (Gumanovskaya et al., Immunology 97, 466-73., 1999); (McKown et al., Arthritis Rheum 42, 1204-8., 1999); (Trentham et al., Science 261, 1727-30., 1993), delivery of insulin to animals and humans with autoimmune diabetes (Pozzilli and Gisella Cavallo, Diabetes Metab Res Rev 16, 306-7., 2000), and delivery of S-antigen to animals and humans with autoimmune uveitis (Nussenblatt et al., Am J Ophthalmol 123, 583-92., 1997). A problem associated with this approach is T-cell unresponsiveness induced by systemic injection of antigen.
Another approach is the attempt to design rational therapeutic strategies for the systemic administration of a peptide antigen based on the specific interaction between the T-cell receptors and peptides bound to MHC molecules. One study using the peptide approach in an animal model of diabetes, resulted in the development of antibody production to the peptide (Hurtenbach, U et al. J Exp. Med 177:1499, 1993). Another approach is the administration of T cell receptor (TCR) peptide immunization. See for example (Vandenbark A A et al., Nature 341:541, 1989). Still another approach is the induction of oral tolerance by ingestion of peptide or protein antigens. See for example (Weiner H L, Immmunol Today, 18:335 1997).
Immune responses are currently altered by delivering proteins, polypeptides, or peptides, alone or in combination with adjuvants (immunostimulatory agents). For example, the hepatitis B virus vaccine contains recombinant hepatitis B virus surface antigen, a non-self antigen, formulated in aluminum hydroxide, which serves as an adjuvant. This vaccine induces an immune response against hepatitis B virus surface antigen to protect against infection. An alternative approach involves delivery of an attenuated, replication deficient, and/or non-pathogenic form of a virus or bacterium, each non-self antigens, to elicit a host protective immune response against the pathogen. For example, the oral polio vaccine is composed of a live attenuated virus, a non-self antigen, which infects cells and replicates in the vaccinated individual to induce effective immunity against polio virus, a foreign or non-self antigen, without causing clinical disease. Alternatively, the inactivated polio vaccine contains an inactivated or ‘killed’ virus that is incapable of infecting or replicating and is administered subcutaneously to induce protective immunity against polio virus.
3. Neurodegenerative Diseases
Neurodegenerative diseases are a broad category of diseases of the central nervous system which are all characterized by a slowly progressive destruction or degeneration of nerve cells (Temlett, Curr Opin Neurol 9, 303-7, 1996); (Dickson, Curr Opin Neurol 14, 423-32, 2001); (Kaye, Neurology 51, S45-52; discussion S65-7, 1998); (Prusiner, Proc Natl Acad Sci U S A 95, 13363-83, 1998); (Cummings et al., Neurology 51, S2-17; discussion S65-7, 1998); (Lin et al., Neuron 24, 499-502, 1999); (Chesebro, Neuron 24, 503-6., 1999); (Ross, Neuron 19, 1147-50., 1997); (Yankner, Neuron 16, 921-32., 1996); (Selkoe, Neuron 6, 487-98., 1991). The degeneration of neurons in the brain or spinal cord leads to devastating permanent clinical symptoms including in some cases profound dementia, abnormal movements, tremor, gait ataxia, or epileptiform activity. Common to nearly all of the neurodegenerative diseases is the progressive dementia which can manifest itself as a complete inability to care for oneself and a total lack of recognition of friends and family.
Another common feature of these diseases is the lack of an effective therapy for any of them. Most of the treatments available today focus on supportive care of the late symptoms and none are directed at the underlying pathophysiologic causes of these diseases. For example, for Parkinson's disease medications are directed at and are usually effective in temporarily controlling the tremor associated with the disease, but no medications are effective in halting the progressive dementia and destruction of neurons within the substantia nigra of the brain (Jankovic, Neurology 55, S2-6, 2000). As another example, in Alzheimer's disease until recently no treatments were available for the progressive dementia that characterizes this disease. Several cholinesterase inhibitors have now been approved for use in Alzheimer's disease (Farlow and Evans, Neurology 51, S36-44; discussion S65-7, 1998) (Hake, Cleve Clin J Med 68, 608-9, 613-4, 616, 2001). These drugs presumably increase the amount of the neurotransmitter acetylcholine available in the brain, leading to improved function of those particular neurons that use acetylcholine as a transmitter. All of these drugs, as a whole, show only miniscule efficacy in clinical trials with the primary endpoint being improvement in cognitive testing. These drugs are also not directed at the primary pathophysiology of Alzheimer's disease, namely the destruction of the cholinergic neurons within the brain. Therefore, no current therapy aimed at the primary pathologic cause exists for any of the neurodegenerative diseases.
The majority of neurodegenerative disease also have in common the finding of aggregated or accumulated substances within the areas of the central nervous system that are most affected by the degenerative process. These abnormal accumulations, that can be found either extra- or intra-cellularly, may contribute to the death and destruction of the relevant neurons. Furthermore, the features and composition of the accumulations are specific for a particular disease. For example, the aggregates in Alzheimer's disease consist of a protein called amyloid beta (Aβ), whereas for Parkinson's disease they are composed of a protein called alpha-synuclein (Dickson, Curr Opin Neurol 14:423-432, 2001); (Cummings et al., Neurology 51, S2-17; discussion S65-7, 1998). The neurodegenerative diseases characterized by the development and accumulation of such aggregates include Alzheimer's disease, Parkinson's disease, Huntington's disease, and prion disease (Yankner, Neuron 16:921-32, 1996); (Ross, Neuron 19:1147-50, 1997) (Chesebro, Neuron 24:503-506, 1999); (Dickson, Curr Opin Neurol 14:423-32, 2001).
4. Polynucleotide Therapy
Gene Therapy. Polynucleotide therapeutics, including naked DNA encoding peptides and/or polypeptides, DNA formulated in precipitation- and transfection-facilitating agents, and viral vectors have been used for “gene therapy.” Gene therapy is the delivery of a polynucleotide to provide expression of a protein or peptide, to replace a defective or absent protein or peptide in the host and/or to augment a desired physiologic function. Gene therapy includes methods that result in the integration of DNA into the genome of an individual for therapeutic purposes. Examples of gene therapy include the delivery of DNA encoding clotting factors for hemophilia, adenosine deaminase for severe combined immunodeficiency, low-density lipoprotein receptor for familial hypercholesterolemia, glucocerebrosidase for Gaucher's disease, α1-antitrypsin for α1-antitrypsin deficiency, α- or β-globin genes for hemoglobinopathies, and chloride channels for cystic fibrosis (Verma and Somia, Nature 389, 239-42, 1997).
DNA immunization to treat infection. In DNA immunization a non-replicating transcription unit can provide the template for the synthesis of proteins or protein segments that induce or provide specific immune responses in the host. Injection of naked DNA promotes vaccination against a variety of microbes and tumors (Robinson and Torres, Semin Immunol 9, 271-83., 1997). DNA vaccines encoding specific proteins, present in viruses (hepatitis B virus, human immunodeficiency virus, rotavirus, and influenza virus), bacteria (mycobacterium tuberculosis), and parasites (Malaria), all non-self antigens, are being developed to prevent and treat these infections (Le et al., Vaccine 18, 1893-901., 2000); (Robinson and Pertmer, Adv Virus Res 55, 1-74, 2000).
DNA to treat neoplasia. DNA vaccines encoding major histocompatibility antigen class I, cytokines (IL-2, IL-12 and IFN-γ), and tumor antigens are being developed to treat neoplasia (Wlazlo and Ertl, Arch Immunol Ther Exp 49:1-11, 2001). For example, viral DNA encoding the B cell immunoglobulin idiotype (antigen binding region) has been administered to eliminate and protect against B cell-lymphomas (Timmerman et al., Blood 97:1370-1377, 2001).
DNA immunization to treat autoimmune disease. Others have described DNA therapies encoding immune molecules to treat autoimmune diseases. Such DNA therapies include DNA encoding the antigen-binding regions of the T cell receptor to alter levels of autoreactive T cells driving the autoimmune response (Waisman et al., Nat Med 2:899-905, 1996) (U.S. Pat. No. 5,939,400). DNA encoding autoantigens were attached to particles and delivered by gene gun to the skin to prevent multiple sclerosis and collagen induced arthritis. (Patent WO 97/46253; Ramshaw et al. Immunol. and Cell Bio. 75:409-413, 1997). DNA encoding adhesion molecules, cytokines (TNFα), chemokines (C-C chemokines), and other immune molecules (Fas-ligand) have been used in animal models of autoimmune disease (Youssef et al., J Clin Invest 106:361-371, 2000); (Wildbaum et al., J Clin Invest 106:671-679, 2000); (Wildbaum et al, J Immunol 165:5860-5866, 2000); (Wildbaum et al., J Immunol 161:6368-7634, 1998); (Youssef et al., J Autoimmun 13:21-9, 1999).
It is an object of the present invention to provide a method of treating or preventing a disease associated with self-protein(s), -polypeptide(s), or -peptide(s) that are present and involved in a non-physiological process in an animal. Another object of this invention is to provide a specific method for treating or preventing autoimmune diseases that does not impair the immune system generally. Still another object of the present invention is to provide a specific method for treating or preventing neurodegenerative diseases. Yet another object of the present invention is to provide a composition for treating or preventing a disease associated with self-protein(s), -polypeptide(s), or -peptide(s) that is present non-physiologically in an animal. Still another object of this invention is to identify self-protein(s), -polypeptide(s), or -peptide(s) that are present non-physiologically and associated with a disease. These and other objects of this invention will be apparent from the specification as a whole.